NORMAL  HISTOLOGY  OF  THE  KIDNEYS, 
LIVER,  HEART,  SPLEEN,  LUNG 

OF  THE 

GUINEA  PIG 

AND 

HISTO-PATHOLOGICAL  CHANGES  OF 
THESE  ORGANS 

IN 

EXPERIMENTALLY  INDUCED  BOTULISM 


BY 

CHARLES  ETHELBURT  RANDOLPH 


THESIS 
FOR  THE 

DEGREE  OF  BACHELOR  OF  SCIENCE 

IN 

AGRICULTURE 

COLLEGE  OF  AGRICULTURE 
UNIVERSITY  OF  ILLINOIS 

1922 


UNIVERSITY  OF  ILLINOIS 


.vhjm..3 19S2.... 

THIS  IS  TO  CERTIFY  THAT  THE  THESIS  PREPARED  UNDER  MY  SUPERVISION  BY 
CHARLES  ETmBURT  RANDOLPH  

ENTITLED.. 9?... 9?.. 
GUINEA  PIG  AND  HISTO-PATHOLOGICAL  CHANGES  OF  THESE  ORGANS  IN  EXPERIMENTALLY 

INDUCED  .BJPUUSM.^ 

IS  APPROVED  BY  ME  AS  FULFILLING  THIS  PART  OF  THE  REQUIREMENTS  FOR  THE 
DEGREE  OF. Ba<^3lpr..of  .Scie^ 


HEAD  OF  DEPARTMENT  OF ... . A$Xml . . Husbandry 


Digitized  by  the  Internet  Archive 
in  2015 


https://archive.org/details/normalhistologyoOOrand 


TABLE  OF  CONTENTS 


Int  roduct  i on — -1 

Fixing  of  Tissues- • ~ — 2 

Embedding  of  Tissues- — • 2 

Cutting  of  Microscopic  Sections • 3 

Staining  of  Sections — - — — --3 

Preparation  of  Delafield's  Hematoxylin---------- — — — ~ — --—3 

Preparation  of  Eosin  Stain--- — 3 

Preparation  of  Xylene-Paraffin  Mixture - - --4 

Preparation  of  Paraffin -Beeswax  Mixture--- 4 

Preparation  of  Japanese  Glycerin-Albumin — — — — 4 

Source  of  Materials----- — — ------4  & 5 


Kidney 

Normal  Histology—————— — — - — — — — — ^ — - — ---5  & 6 

Histo-Pathological  Changes-- — — ----6  Sc  7 

Liver 

Normal  Histology------ — — — — •--  — -7  & 8 

Histo-Pathological  Changes-—— — — >-8  & 9 

Heart 

Normal  Histology* — ------- — — - — 9 & 10 

Histo-Pathological  Changes — ------10  & 11 

Spleen 

Normal  Histology — — ------- — — — ---11  & 12 

Histo-Pathological  Changes---- — — — >12 

Lung 

Normal  Hist o logy — — — — — — « — - — 13 

Histo-Pathological  Changes------ — ————-13  & 14 

Summary  of  Pathological  Changes--—- — — — — =-14 

Bibliography———— — — - — — « 15 

Diagnosis  Charts- — — — — — —16  to  20 

inclusive 


NORMAL  HISTOLOGY  OF  THE  KIDNEYS, 
LIVER,  HEART,  SPLEEN,  LUNG 
OF  THE 
GUINEA  PIG 

AND 

HISTO-PATHOLOGICAL  CHANGES  OF 
THESE  ORGANS 
IN 

EXPERIMENTALLY  INDUCED  BOTULISM 


************ 

The  sporadic  outbreaks  of  botulism  in  animals  reported  in  recent  years 
has  suggested  the  desirability  of  accurate  diagnostic  methods.  In  a peracute 
attack  of  the  atypical  type  of  this  disease  the  symptoms  are  not  pathognomonic, 
and  are  probably  frequently  confused  with  other  diseases.  The  presence  of 
the  spore  in  the  intestinal  tract,  or  in  the  internal  oigans  as  an  agonal  in- 
vasion following  death,  is  not  irrefutable  diagnostic  evidence  of  botulism. 

The  microscopic  changes  in  the  tissues  of  the  guinea  pig,  after  experimentally 
inducing  botulism,  have  been  studied  with  the  view  of  determining,  if  possible, 
the  value  of  histo-pathological  changes  in  the  kidneys,  liver,  heart,  spleen 
and  lungs.  In  the  following  paper  a brief  study  of  the  histology  of  the  normal 
guinea  pig  and  a study  of  tissues  obtained  from  cases  of  experimentally  induced 


botulism  are  recorded. 


- 2 - 


METHOD  OF  FIXING,  EMBEDDING,  CUTTING  AND  STAINING  OF  TISSUES 
Fixing  and  Embedding  Tissues; 

It  is  essential  in  the  sectioning  of  body  tissues  to  retain  the  normal 
relationship  of  the  cells  of  the  organ  as  present  in  the  individual.  Post  mor- 
tem changes  take  place  rapidly  so  that  the  tissues  should  be  placed  in  a fixing 
fluid  (10  Formalin)  as  soon  as  possible.  After  the  fixation  is  complete  the 
tissues  are  washed  and  later  dehydrated  by  placing  them  in  a series  of  solutions 
of  increasing  concentrations  of  alcohol. 

Prior  to  the  process  of  embedding,  the  tissues  are  cleared  in  xylene,  and 
after  being  completely  infiltrated  with  hot  paraffin,  they  are  embedded  in  blocks 
of  the  same  material.  The  following  technic  was  employed  in  fixing,  embedding, 
cutting  and  staining  the  preparations.  The  tissues  for  sectioning  purposes  were 
cut  about  two  centimeters  square  and  after  being  washed  free  of  excess  blood 
they  were  placed  in  a volume  of  the  1C  $ formalin  solution  about  fifty  times  the 
volume  of  the  tissue. 

Fixing  and  Embedding  Technic: 

10  % Formalin  in  physiologic  salt  solution  for  24  hrs. 

Wash  for  24  hrs. 

70  $ Alcohol  24  hrs  (or  longer). 

S5  $ Alcohol  24  hrs. 

Absolute  Alcohol  24  hrs. 

Xylene  2 hrs. 

Xylene -Paraffin  Mixture* **--2  hrs)_=jn  oven  at  58°C. 

Paraffin-Beeswax  Mixture ***-4  hrs) 

Embed  in  Paraffin-Beeswax  Mixture 

The  tissues  are  embedded  in  paper  boats,  which  are  of  common  white  paper  about 

eight  centimeters  long  and  three  centimeters  wide.  Each  boat  is  placed  upon  a 

cold  surface  and  filled  half  full  of  the  warm  paraffin-beeswax  mixture.  Then 

with  warm  forceps  the  square  of  tissue  is  placed  in  the  boat,  the  surface  to  be 

cut  being  embedded  down.  The  filling  of  the  boat  is  completed  with  the  warm 

paraffin.  This  second  pouring  will  drive  out  bubbles  of  air  which  adhere  to  the 

bottom.  The  boat  is  then  floated  in  cold  water  to  speed  the  solidification  of 
the  paraffin  and  drive  out  air. 

^**See  preparation  on  page  4. 


■ 


- 3 - 


Cutting  of  Microscopic  Sections: 

The  paper  boats  are  stripped  from  the  hardened  paraffin  and  the  blocks 
trimmed,  taking  care  that  the  edges  of  the  cutting  surface  are  parallel  to  in- 
sure ribboning  of  the  sections.  A horizontal  sliding  microtome  was  used  in  this 
project.  Four  microns  was  the  usual  thickness  of  the  sections,  but  with  certain 
blocks  it  was  only  possible  to  get  good  sections  at  five  or  six  microns.  The  cut 
sections  were  floated  on  warm  water  and  mounted  on  a slide  by  putting  the  slide 

underneath  the  floating  section  and  raising  it  out  of  the  water.  The  sections 

* * 

were  held  to  the  slide  by  a Japanese  glycerine-albumin  mixture  previously  rubbed 
on  the  slide  and  heated  until  a steam  began  to  arise  from  the  surface.  The 
mounted  sections  were  allowed  to  dry  in  an  oven  at  58°C  for  12  hours. 

Staining  of  the  Sections: 

Hematoxylin  and  eosin  ware  used  for  staining  all  tissues.  The  hematox- 
ylin stains  the  nuclei  purple  and  the  cytoplasm  takes  the  eosin  or  acid  stain. 
Preparation  of  Delafield's  Hematoxylin. 

Hematoxylin  crystals--- A grams 

Alcohol  (absolute) — 25  cc. 

Saturated  solution  of 

ammonium  alum-400  cc. 

The  hematoxylin  crystals  are  dissolved  in  the  absolute  alcohol.  To 

this  solution  400  cc  of  alum  solution  was  added.  The  mixture  was  exposed  to 

the  light  (sunlight  if  possible)  in  an  unstoppered  bottle  for  3 to  4 days. 

After  filtration  the  following  were  added: 

Glycerine----- — - — ---100  cc 

Alcohol  (absolute) — — 100  cc 

This  solution  was  allowed  to  stand  in  the  light  until  the  color  was  sufficiently 

dark,  then  filtered  and  kept  in  a tightly  stoppered  bottle.  The  solution  keeps 

well  and  is  satisfactory  as  long  as  it  retains  a purplish  tinge. 

Preparation  of  Eosin  Stain: 

Eosin-- — --1  gram 

Water- — — 100  cc 

Absolute  alcohol 15  cc 

The  eosin  was  dissolved  in  the  distilled  water,  the  alcohol  added,  and  filtered. 
♦♦Preparation  on  page  4.  


- 4 - 


The  staining  was  accomplished  by  the  following  technic: 

Xylene — 3 min. 

Absolute  Alcohol ------ -2  min. 

95$  Alcohol-- - 2 min. 

60$  Alcohol — ---■ — 2 min. 

Distilled  HgO >• 2 min. 

Hematoxylin-- — 6to30  min. 

Tap  water — -2  min. 

Distilled  HgO------ 2 min. 

Eosin ——2  to  3 min. 

Tap  water-— -2  min. 

Distilled  HpO— — 2 min. 

70$  Alcohol-  — ------- 1 min. 

95$  Alcohol ------ -----3  min. 

Absolute  Alcohol-— --4  min. 

All  excess  staining  matter  on  the  slide  around  the  tissue  was  removed  with  a 
cloth. 

Xylene — - — --—3  min. 

The  surplus  xylene  was  cleaned  off  and  a drop  of  Canadian  balsam  added  to  the 
section  over  which  a cover  slip  was  then  gently  placed  taking  care  to  exclude 
all  air  bubbles.  A light  weight  was  placed  on  the  cover  slip  to  hold  it  in 
place  until  the  balsam  dried.  The  slides  were  sat  aside  to  allow  the  balsam  to 
dry  and  seal  the  cover  slip. 

Preparation  of  Xylene-Paraff in  Mixture: 

1 part  56  JC  paraffin 

1 part  46° C paraffin 

2 parts  xylene 

Slowly  melt  the  paraffin  and  add  the  xylene  to  the  liquid  paraffin. 

Preparation  of  Paraffin-Beeswax  Mixture 

Paraffin  850  grams. 

Stearin  100  grams 
Beeswax  50  grams 

This  mixture  is  kept  in  the  oven  at  56°C. 

Preparation  of  Japanese  Glycerin-Albumin, 

White  of  Egg) 

) — -Equal  parts 

Glycerine  ) 

Beat  mixture  and  filter.  Add  a few  drops  of  formalin  as  a preservative. 

SOURCE  OF  MATEEIALS 

The  normal  tissue  specimens  which  were  used  for  comparative  purposes 


. 


- 5 - 


were  obtained  by  destroying  a healthy  guinea  pig  and  immediately  removing  the 
fresh  organa.  The  guinea  pigs  furnishing  the  pathological  tissue  specimens  were 
originally  used  in  botulinua  typing  tests.  A majority  of  the  guinea  pigs  (13) 
died  in  a few  hours  ( 12-30  hrs)  following  an  acute  attack  of  botulism,  while 
two  pigs  were  destroyed  in  the  agonal  stages  of  the  disease  in  order  to  avoid 
the  development  of  post-mortem  changes. 

KIDNEYS 

Normal  Histology: 

The  kidneys  are  glandular  organs  located  on  each  side  of  the  dorsomed- 
ian  line.  They  are  invested  in  a fibrous  capsule  which  blends  into  the  ureter 
at  the  hilum.  The  organ  is  divided  into  four  regions,  the  cortex,  medulla,  in- 
t . ate  zone,  and  pelvis.  Extending  into  the  sinus  at  the  hilum  are  the  re- 

r .1  papillae,  the  projections  of  the  renal  pyramids.  These 
renal  pyramids  extend  back  thru  the  medulla  and  have  their 
cu\.,w..tBwKwi  base  towards  the  cortex.  Between  the  pyramids  are  found 

-UftW 

^ the  columns  of  Bert  ini  which  exter.d  to  the  walls  of  the 

;i  1 nus . 

On  the  examination  of  the  cortex  under  low  power  magnification  it  is  seen 
that  it  is  divided  into  alternating  light  and  dark  areas,  the  medullary  rays  and 


*4*^ The  structure  of  a renal  secretory  unit  is  composed  of  three 


general  parts  namely,  the  glome:  ulus, the  convoluted  urinfer- 


''‘•~-T«xin-vrU.vA t tubules  and  the  collecting  tubules  or  duct  system.  As 

U.oU 

. ' shown  in  the  figure  the  uriniferous  tubules  begin  in  a great 

ly  expanded  blind  extremity,  the  capsule.  This  capsule  sur- 

u«^Wt  rounds  the  tuft  of  an  artery  forming  the  Malpighian  body. 

Th-.  • >ule  coming  from  the  glomerulus  takes  a very  tortuous  and  twisted  course 
forming  the  proximal  convoluted  tubule.  It  then  extends  into  the  medullary  rays 
and  follows  a straight  course  towards  the  papillae,  as  the  descending  limb  of 
Henle.  When  well  into  the  medullary  region  it  turns  upon  itself,  the  loop  of 


fm 


- 6 - 

Henle,  and  returns  to  the  renal  pyramid  and  the  medullary  rays  as  the  ascending 
limb  of  Henle.  It  again  enters  the  labyrinth  and  repeats  its  tortuous  convoluted 
course,  forming  the  distal  convoluted  tubule.  Later  it  reenters  the  medullary 
rays,  and  joining  one  of  the  collecting  ducts,  it  finds  its  way  into  the  renal 
pyramid  and  out  through  the  papillae  into  the  sinus. 

The  structure  of  the  urinary  tubules  is  very  simple,  consisting  of  a 
single  layer  of  cells  supported  by  a basement  membrane.  The  glomerular  portion 
eliminates  the  water  and  inorganic  constituents,  while  the  organic  matter  is 
eliminated  thru  the  tubules,  by  this  arrangement  the  tubules  are  continually 
washed  by  the  liquid  which  carries  the  secretory  products  down  the  tubule.  The 
urine  collects  in  the  sinus  which  has  an  outlet  thru  the  ureter. 

KIDL'EY 

Histo-Pathological  Changes  2 

Parenchymatous  degeneration  of  the  urinary  tubules  was  the  most  constant 
characteristic  pathological  change  found  in  the  kidneys.  The  degree  of  degener- 
ation varied  from  a very  severe  one  bordering  on  necrosis  to  a very  mild  degen- 
erative disturbance.  In  the  majority  of  the  cases  the  parenchymatous  degenera- 
tion was  marked. 


Kidney — Histopathological  Changes 


1 


7 


Passive  congestion  existed  in  over  80$  of  the  cases.  Over  half  of  the 
series  (.13  cases)  presented  advanced  passive  congestion;  the  congestion  backing 
up  to  involve  the  glomeruli.  The  passive  congestion  which  involved  the  glomeru- 
li was  classified  as  severe,  while  the  less  severe  cases  were  rated  as  marked 
and  slight.  Ten  percent  of  the  kidneys  were  markedly  congested,  while  30$  were 
only  slightly  congested,  A final  10$  showed  no  passive  congestion. 

There  were  also  other  changes  present  in  a few  instances.  A few  hemorr- 
hages were  found  in  three  cases.  One  kidney  showed  a beginning  necrosis  of  the 
urinary  tubules  around  the  hemorrhagic  area.  Another  organ  also  presented  a few 
small  areas  of  necrosis  and  one  showed  a slight  edema  in  the  cortex  region. 

In  general  the  most  constant  changes  were  a marked  parenchymatous  degen- 
eration and  a marked  passive  congestion. 


LIVER 


The  Normal  Histology: 

The  liver,  the  largest  gland  in  the  body,  consists  of  very  delicate  glan- 
dular tissue  disposed  around  ramifications  of  the  portal  vein.  It  is  covered 
with  a capsule  of  fibrous  tissue  giving  off  trabeculae  which  extend  into  the  in- 
terior. The  position  of  the  liver  in  the  body  cavity  is 
immediately  below  the  diaphragm  chiefly  on  the  right  side 
with  the  smaller  lobe  extending  into  the  left  side  of  the 
cavity.  Grossly  the  liver  is  divided  into  two  large  lobes. 
Each  of  these  lobes  is  subdivided  by  the  interlobular 
tissue  into  many  lobules.  Centrally  located  in  each  lob- 
ule, the  central  vein  receives  the  capillary  branches  of 

The  hepatic  cells,  the  smallest  unit  of  structure  of  the  liver,  are  poly- 
gonal in  shape  and  contain  a comparatively  large  nucleus.  The  cytoplasm  is 
finely  granular,  and  the  nucleus  contains  a reticularly  arranged  mass  of  chro- 
matin material.  The  liver  cells  are  placed  in  rows  or  cords  projecting  from  the 


the  portal  vein. 


■ 


. 


- 6 - 

central  vein-like  spokes  in  a wheel. 

The  biliary  sinuses  between  the  rows  of  cells  which  collect  the  secreted 
bile  from  the  liver  cells  have  no  definite  vessel  walls  but  their  course  follows 
thru  trough-like  depressions  in  the  walls  of  the  liver  cells.  The  larger  bile 
vessels  have  true  walls  and  are  usually  found  in  close  proximity  to  a hepatic 
vein  and  a hepatic  artery  forming  the  so-called  hepatic  trinity. 

Two  blood  supplies,  namely,  the  functional  and  the  nutritional,  are  found 
in  the  liver.  The  functional  supply  comes  from  the  portal  vein.  After  leaving 
the  portal  vein  the  blood  flows  into  the  interlobular  branches  of  the  vein  which 
carry  it  thruout  the  liver  and  brings  it  to  the  individual  lobules.  These  in- 
terlobular veins  then  give  off  capillary  ramifications  which  extend  thruout  the 
lobule  and  bring  the  blood  in  intimate  contact  with  the  hepatic  cells.  Later 
the  capillaries  collect  at  the  central  vein  and  conduct  the  functional  blood  to 
the  sublcbular  branch  of  the  hepatic  vein  and  finally  back  to  the  heart.  The 
amount  of  interstitial  tissue  varies  in  different  species.  In  swine  the  lobules 
are  very  distinct  due  to  the  large  amount  of  interlobular  connective  tissue. 

The  nutritional  blood  is  derived  from  the  hepatic  artery  and  follows  a 
course  somewhat  similar  to  the  functional  blood.  Later  the  two  unite  and  both 
return  to  the  heart  by  way  of  the  same  vessels. 

LIVES 

tiisto-Pathological  Changes: 

Passive  congestion,  tho  not  present  in  all  the  livers  was  characteristic 
of  them  in  general.  Two  livers  were  severely  congested,  two  others  showed  mark- 
ed passive  congestion,  and  ten  were  only  slightly  congested  while  five  presented 
no  noticeable  congestion.  There  were  no  hemorrhages  in  any  of  this  series  (19 
cases) . 

The  most  characteristic  pathological  changes  in  the  livers  were  fatty 
degeneration  and  fatty  infiltration.  Only  two  cases  could  be  classed  as  a pure 
fatty  infiltration  while  fifteen  cases  showed  fatty  degeneration  with  some  fatty 


• ■ 


_ 9 - 


infiltration  in  three  cases.  It  was  interesting  to  note  that  there  were  only 
two  cases  which  were  free  of  pathological  fat. 


L i ver — Hi s t o-Pat h c 1 c gi ca 1 Change  s 

Sixteen  of  the  guinea  pigs  showed  marked  parenchymatous  degeneration  of 
the  liver.  Most  of  the  parenchymatous  degeneration  was  present  with  fatty  degen- 
eration* while  there  were  only  two  mild  cases  of  parenchymatous  degeneration  in 
livers  which  did  not  show  fatty  degeneration.  All  cases  showed  either  a paren- 
chymatous or  fatty  degeneration.  A slight  cellular  hepatitis  was  found  in  four 
cases. 

Passive  congestion,  marked  fatty  degeneration,  and  parenchymatous  degen- 
eration were  the  three  most  characteristic  pathological  changes  in  the  liver. 


HE  APT 

Normal  Histology: 

Histologically  the  heart  is  composed  of  three  layers  of  tissue,  the  epi- 
cardium,  myocardium  and  endocardium.  The  endocardium  is  composed  of  a single 
layer  of  endothelial  plate-like  cells  and  underlying  connective  tissue  which  is 
quite  rich  in  elastic  fibers  with  a small  quantity  of  involuntary  muscle.  This 

layer  is  composed  chiefly  of  muscle  fibers  which  are  not  organized  into  bundles. 


10 


but  form  a very  complex  system  of  branching  and  intermingling  fibers  or  intercal- 
ated discs.  Between  the  branching  cardiac  muscle  structure  the  connective  tissue 
aids  in  binding  the  fibers  together.  There  is  no  well  defined  endomysium  as  in 
voluntary  muscle. 

The  outer  layer  of  the  heart,  the  epicardium,  consists  of  a single  layer 
of  endothelial  cells  having  a substratum  of  f ibro-elast ic  tissue.  The  epicardi- 
um completely  covers  the  heart  and  blends  into  the  adventitia  of  the  great  veins. 

HEART 

Histo-Pathclogical  Changes: 

The  only  constant  pathological  change  in  the  heart  was  a parenchymatous 
degeneration.  Most  of  the  heart  series  (16  cases)  were  only  slightly  degenerated, 
while  two  showed  marked  degeneration,  and  one  severe  degeneration. 


Heart--Histo-Pathological  Changes 

The  striations  of  the  muscle  fibers  were  not  visible  due  tc  the  parenchymatous 
degeneration.  Congested  coronary  vessels  were  found  in  six  specimens  of  this 
organ  and  one  showed  a passive  congestion  of  the  capillaries  of  the  cardiac  mus- 
cle. Another  change  present  in  three  of  the  hearts,  which  might  have  been  due 


11 


to  the  method  of  fixation,  was  a separation  of  the  muscle  fibers  at  the  cement 
lines.  The  fibers  of  one  of  the  hearts  appeared  as  tho  they  were  shrunken. 

Summarizing  the  series  of  the  heart  it  was  found  that  parenchymatous  de- 
generation was  characteristic  thruout  and  a congestion  of  the  coronary  vessels  in 
a few  cases. 


THE  SPLEEN 


Normal  Histology: 

The  spleen  may  be  considered  as  a huge  hamolymph  node,  possessing  the 
functions  of  producing  lymphocytes  and  of  destroying  erythrocytes.  It  is  dark 
red  in  color  due  to  the  fact  that  it  contains  a large  quantity  of  blood.  This 
organ  is  located  on  the  left  side  of  the  abdominal  wall  between  the  stomach  and 
the  diaphragm.  The  spleen  is  invested  in  a capsule  composed  of  dense  elastic 
tissue  with  a few  scattered  fibers  of  involuntary  muscle.  Extending  from  the 
capsule  into  the  spleen  are  the  trabeculae  which  branch  into  a delicate  frame- 
s^...X^So\e.  work  giving  support  and  a degree  of  rigidity  to  the  organ. 
syW&lL  Histologically  the  spleen  is  composed  of  splenic  lobules. 

These  are  imperfectly  defined  by  the  interlobular  trabecu- 

-'Kay  ecu  la 

if  lae.  Branching  from  these  interlobular  trabeculae  the  in- 


■ T'--  ' ' • 


tralcbular  trabeculae  divide  the  3plenic  lobule  into  about 


ten  primary  compartments.  The  space  between  this  frame-work  of  trabeculae  is 
filled  by  splenic  pulp  and  lymph  follicles.  The  splenic  pulp  is  composed  of 
essentially  four  elements  namely,  lymphocytes,  leucocytes,  erythrocytes  and  the 
large  phagocytic  cells. 

The  blood  enters  at  the  hilum  and  after  traversing  in  the  hilum  connec- 
tive tissue  for  a short  distance  the  arteries  breakup  into  branches  which  supply 
individual  splenic  lobules.  Prolongations  of  the  connective  tissue  of  the  trab- 
eculae follow  the  arterial  branches  into  the  pulp,  thus  surrounding  them  with  an 
extra  fibrous  envelope.  Between  this  envelope  and  the  artery  wall  local  accumu- 
lations of  lymphocytes  occur  giving  rise  to  a spherical  or  fusiform  mass  of 
lymphoid  tissue,  the  Malpighian  bodies.  The  functional  blood  supply  after  con- 


. 


12 


tinuing  in  vessels  for  a time,  after  entering  at  the  hilurn,  a part  leaves  the 
vessels  and  flows  freely  thru  the  splenic  pulp,  while  another  part  continues  thru 
thin  walled  venous  sinuses.  There  is  a slow  movement  of  the  blood  thru  the  spleer 
after  which  it  is  again  collected  in  vessels  and  carried  away.  The  nutritional 
blood  supply  on  the  other  hand  is  conducted  thruout  its  course  in  closed  vessels. 

SPLEEN 

Histo-Pathological  Changes: 

Follicular  hyperplasia  was  the  outstanding  feature  of  over  85$  of  the 
spleens.  Most  of  the  cases,  however,  were  mild  and  only  two  were  marked.  Con- 
gestion was  present  in  65$  of  the  cases.  In  one  spleen  there  was  an  increased 
number  of  leucocytes  while  three  others  had  eosinophilic  infiltrations.  A mild 
splenitis  was  found  in  one  specimen. 


Spleen--Histo -Pathological  Changes 

The  increase  in  the  size  of  the  splenic  follicles  and  the  congestion  were 
the  most  outstanding  changes. 


- 


. 

13 


THE  LUNGS 


Normal  Histology: 

The  lungs,  the  respiratory  organs  of  the  body,  are  composed  of  thin 
walled  air  sacs  surrounded  by  thin  walled  capillaries  which  allow  the  air  to 
come  in  close  contact  with  the  blood  and  permit  a gaseous  exchange. 

On  examining  the  outer  surface  of  the  lung  it  is  seen  that  it  is  divided 
into  polygonal  areas,  or  lobules  which  are  the  unit  of  structure  of  the  lung, 
beginning  at  the  trachea  the  air  passages  divide  as  seen  in  the  figure  below. 

The  trachea  extends  well  down  into  the  thorax  and  then  divides  into  two  bronchi, 
one  going  to  each  lung.  The  bronchus  on  entering  the  lung  begins  to  divide  and 
subdivide.  The  branches  are  called  bronchioles.  The  larger  bronchioles  have 
cartilaginous  rings  as  in  the  walls  of  the  main  bronchi.  The  medium  sized 
bronchioles  contain  only  scattered  plates  of  cartilage.  These  smaller  bronchi- 
oles divide  and  finally  end  in  terminal  bronchioles.  The  terminal  bronchioles 
are  connected  with  the  alveoli  or  air  sacs  by  means  of  the  atria. 

On  cross  section  the  lung  appears  as  shown  in  figure  below.  The  walls 
of  the  air  sacs  are  composed  of  flattened  spherical  cells.  The  blood  supply 
follows  the  divisions  of  the  bronchioles  and  extends  into  the  interstitial  tissue 
between  the  air  sacs. 


The  walls  of  the  bronchioles  are  composed  of  three  parts, 
.namely,  the  fibrous  tunic  which  contains  the  cartilaginous 



' fllveal‘'  rings,  the  central  layer  of  smooth  muscular  and  connective 

TW—' ' I tissues,  and  the  mucous  membrane  lining  the  inner  surface 

of  the  lumen. 


THE  LUNGS 

Histo-Pathological  Changes: 

Passive  congestion  of  the  lunsrs  was  present  in  all  of  the  cases  but  one. 
Three  of  the  lungs  apparently  showed  hemorrhages,  which  might  be  the  result  of 
agonal  struggling  in  those  few  cases  destroyed  by  concussion,  Seren  of  the  lungs 
were  severely  congested  and  six  only  slightly  congested.  One  lung  was  normal. 


' 


■ 


14 


Lung- -His to -Pathological  Changes 

A slight  catarrhal  exudate  was  present  on  the  edges  of  the  air  sacs  in 
one  lung  and  another  had  a catarrhal  bronchial  exudate.  Five  lungs  presenting 
advanced  passive  congestion  also  showed  sub-pleural  edema. 

SUMMARY 

The  most  consistent  histo-pathological  change  in  the  kidney,  liver, 
heart,  spleen  and  lung  of  guinea* pigs  suffering  from  an  acute  form  of  botulinu3 
poisoning  was  a passive  congestion,  with  parenchymatous  degeneration  in  varying 
degrees.  The  later  in  a few  instances  bordered  on  necrosis.  Occasionally  be- 
ginning inflammatory  changes  were  encountered,  but  in  the  series  as  a whole  this 
was  not  characteristic.  Similar  changes  may  be  observed  in  other  toxemic  dis- 
eases, and  are  of  little  or  no  value  in  rendering  a differential  diagnosis.  The 
acute  character  of  the  disease  in  the  guinea  pigs  that  furnished  the  tissues  for. 
these  studies  may  not  be  typical  of  changes  in  a chronic  form  of  botuiinus  in- 
t oxicat ion, 

***************** 


• : 


. - • 


- lb 


BIBLIOGRAPHY' 

Textbooks  in  Histology 

Normal  Histology' — Pier  sol 

Essentials  of  Histology----- — ---LeRoy 

A Compend  of  Histology — • — Eadasch 

Textbooks  in  Pathology 

Special  Pathologic  Studies(Two  volumes) Durck 

General  Pathology-- -Durck 

Principles  of  Pathologic  Histology -Mallory 

Manual  of  Pathology — McConnel 

References  in  Botulinus  Poisoning 

McCaskey,  G.W. , 19 IS, Bacillus  Botulinus  Poisoning. 

Am. Jour. Mel. Sc. July  1919  No. l Vol.clviii  p 5? 

Dick  son, Ernest  C. . 19 13, Botulism, Archives  of  Internal  Medicine 
Oct . 1913, Vo 1. XXII, pp  483-495. 

Dickson, Ernest  C. , 19 13, Botulism, Monographs  of  the  Rockefeller 
Institute  for  Internal  Medicine, No. 8 July  31,1913. 


. 


16 


DIAGH0S1S  CHART  OF  KiDHiL'YS 

Ho. 

Blood. 

Cloudy  Swelling  Remarks 

8-1 -A 

Congestion-Localized  hem* 
Slight  hem. in  glomeruli. 

C.5**in  varying  degrees.  Slight  edema 
Some  border  on  necrosis 

9-1 -A 

Passive  Congestion. 
Slight  cong.in  glom# 

Marked  c.s.of  T.***  Collection  of  red 

cells  under  cap- 
suled! ight  nec- 
rosis at  hem. 

13-b 

Severe  passive  cong. 

Marked  c.s.of  T. 

14-c 

Passive  Congestion 

Marked  c.s.of  T. 

15-C 

Very  severe  cong. 

Marked  c.s.of  T . 

lo-c 

Slight  pass.cong. 

Mild  c .s.of  T. 

1 7-2-A 

Severe  pass.cong. 

Mild  c.s.of  T. 

18-1- A 

Severe  pass.cong. 

Marked  c.s.of  T.  Areas  of  necrosis 

19-A 

Severe  pass.cong. 

C.s.of  T. 

21-1-ABeginning  pass.cong. 

Slight  c.s.of  T. 

23-  2- A 

Severe  pass.cong. 

Severe  c ,s  .of  T. 

24-A 

Marked  c.s.of  T. 

25-A 

Severe  pass.cong. 

Slight  c.s.of  T. 

2b-C 

Slight  c.s.of  T. 

27-s 

Slight  c.s.oi  x. 

2Z-a 

Marked  pass.cong. 

Marked  c.s.of  T. 

29 -A 

Slight  pass.cong. 

Slight  c.s.of  T 

30-D 

Slight  pas. cong. 

Slight  c . s .of  T . 

31 -A 

Marked  pas. cong. 

Slight  c.s.of  T. 

*Hemorrhage  C. s.**uloudy  Swelling  of  Urinary  Tubules 
fGlomeruli  . 

17 


No  . 

Blood 

DIAGNOSIS  CHART 
Cloudy  Swelling 

OF  THE  LIVERS 
Degenerations 

Remarks . 

8 

Severe  pass 

Slight  c.s.* 

Fatty  infilt .& 

Slight  cell- 

13-A 

cong. 

Pass. cong. 

Severe  c.s. 

degene rat  ion 
Fatty  infilt  & 

ular  hepatitis. 

14-D 

Slight  p.c"* 

Severe  c.s. 

deg.  (slight ) 
Slight  fatty  deg. 

15-B 

Marked  p.c. 

Slight  c.s. 

Slight  fatty  deg. 

16-B 

Slight  p.c. 

C.s. 

Severe  fatty  deg. 

Slight  cellular 

17-2-C 

Mild  p.c. 

C.s. 

Fatty  deg.&  Infilt. 

hepatitis. 

18-l-C 

Slight  p.c. 

Marked  c.s. 

Fatty  deg. 

Deg. near  necrosis 

19-C 

Slight  P.c. 

Marked  fatty  deg. 

Slight  cellular 

21 -C 
23-l-C 

Severe  p.c. 

Marked  c.s. 

Marked  fatty  deg. 
Marked  fatty  deg. 

hepatitis 

24-C 

Slight  c.s. 

Severe  fatty  deg. 

Slight  cellular 

25-D 

Marked  p.c. 

Marked  c.s. 

Marked  fatty  deg. 

hepatitis 

26- B 

27- C 

Mild  p.c. 

Mild  c.s. 

Mild  fatty  deg. 
Mo  fatty  deg. 

28- E 

29- A 

30- B 

Slight  p.c. 
Slight  p.c. 

Marked  c.s. 

Slight  c.s. 
Slight  c.s. 

Fatty  infilt. on 
piripheral  areas 

Marked  fatty  deg. 

31-D 

Slight  p.c. 

Marked  c.s. 

Slight  fatty  deg. 

c.s*cloudy  swelling 

♦♦Passive 

Congestion 

. . 


. . 

. . 


No. 

13- 0- 

14- B 

15- A 

17- 1- 

18- 1- 
21 -B 

23- 1- 

24- B 
2 i>-B 

26- E 

27- A 

28- D 

29- D 

30- C 

31- C 


DIAGNOSIS  CHART  OF  THE  HEARTS 


Blood 


Cloudy  Swelling 


Remarks 


Cong. of  coronciry 
vessels 

i Pass.cong.of  cap- 
illaries of  car- 
diac muscle . 


{ 


Coronary  vessels 
congested 

Cong. of  coronary 
vessels . 


No  congestion 


Cong. of  coronary 
vessels 


Striations  not  visible  due 
to  c.s.* 

Marked  c.s. All  striations 
gone 

Severe  c.s. 


Marked  c.s. 
Striations  gone 


Slight  c.s. 
Striations  gone 


Mild  c.s. 

Very  slight  c.s. 


Striations  visible  in  part 
of  the  fibers 

Slight  c.s. 


Very  mild  c.s. 
Slight  c.s. 

Si ight  c.s. 

Veyy  little  c.s. 
Slight  c.s. 
Slight  c.s. 


Slight  separation  at 
cement  lines°Fibers 
shrunken . 


Slight  separation  at 
cement  lines. 

Separation  at  cement 
I ines . 


*Cloudy  Swelling 


' 


■ 


DIAGNOSIS  CHART  OF  THE  SPLEENS 


No. 

Blood 

Follicular  Hyperplasia 

Remarks 

9-i-c 

General  cong. 
Increase  in  leuk* 

Mild  f.h.** 

13-F 

Congestion 

14-F 

f .h. 

15-2-A 

Slight  f.h. 

16-E 

No  congestion 

Slight  f.h. 

17-l-B 

Blight  f.h. 

18-2-B  Mild  congestion 

19-E 

Slight  cong. 

Slight  f.h. 

21-3-B 

Mild  interstitial 
splinitis 

23-2-C 

Slight  cong. 

f.h. 

24- F 

Congestion 

f.h. 

26-F 

Focal  collections  of 
eosinphiles . 

27-F 

f.h. 

Eosinphilic  infiltration 

28-F 

Very  slight  cong. 

Slight  f.h. 

Increase  of  eosinphiles 

29-F 

Very  slight  cong. 

Marked  f.h. 

31-E 

Mild  f.h. 

♦Leucocytes  ♦♦Follicular  Hyperplasia 

, 


- 20  - 


DIAGNOSIS  CHARTS  OF  THE  LUNGS 

No. 

Blood  Exudate 

Remarks 

8-1 -D 

Passive  cong*  Slight  catarrhal 

the  edges  of  air 

exudate  on 
sac  3 . 

9-2-d 

Very  severe  passive 
congestion 

Mild  compensatory 
emphysema 

13-E 

Slight  congestion 

14-A 

Severe  cong. in  most 
areas°Hem .over  a large 
area. 

Sub-Pleura  edema 
A farw  eosinphiles. 

15-D 

Slight  passive  cong. 

Suggestive  of  emph 
ysema. 

16-A 

Slight  passive  cong. 

17-1-E 

Severe  passive  cong. 

Slight  sub-pleura 
edema . 

18-1-D 

Severe  passive  cong. 

19-D 

Slight  passive  cong. 

21 -2- A 

Hypostatic  cong. 

Desquamation  of  epth. 
cells  of  air  sacs. 

23-1 -B 

Very  severe  cong. 

Ed  ema- D e s quarna  t i 0 n 
of  alveloir  cells. 

24-1-E 

Marked  congestion 

Slight  edema 

25-C 

Very  severe  cong. 

26 -A 

Slight  congestion 

27- E 

Normal 

28-A 

Hemorrhage 

29-E 

Very  marked  cong.  Slight  exudate  in  bronchi 

30-A 

Slight  passive  cong. 

31-B 

Hemorrhage 

Edema- Blood  in 
bronchi . 

♦Congestion 

